Inhomogeneous translational diffusion of monoclonal antibodies on phospholipid Langmuir-Blodgett films
Abstract
The translational mobility of fluorescent-labeled monoclonal antibodies specifically bound to supported phospholipid bilayers containing hapten-conjugated phospholipids has been measured as a function of the surface concentration of bound antibodies using fluorescence recovery after photobleaching. Fluorescence recovery curves are fit well by a model that assumes the presence of two populations of antibodies with different lateral diffusion coefficients. The larger diffusion coefficient equals 3.5 x 10(-9) cm2/s, the smaller diffusion coefficient ranges from 1.5 x 10(-9) cm2/s to 2.5 x 10(-10) cm2/s, and the fractional fluorescence recovery associated with the smaller coefficient increases from approximately 0 to approximately 0.7 with increasing concentration of bound antibody. These results suggest that complexes of haptenated phospholipids and antibodies in phospholipid Langmuir-Blodgett films form clusters or domains in a concentration-dependent fashion.
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Selected References
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- Andries C., Guedens W., Clauwaert J., Geerts H. Photon and fluorescence correlation spectroscopy and light scattering of eye-lens proteins at moderate concentrations. Biophys J. 1983 Sep;43(3):345–354. doi: 10.1016/S0006-3495(83)84358-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Balakrishnan K., Hsu F. J., Cooper A. D., McConnell H. M. Lipid hapten containing membrane targets can trigger specific immunoglobulin E-dependent degranulation of rat basophil leukemia cells. J Biol Chem. 1982 Jun 10;257(11):6427–6433. [PubMed] [Google Scholar]
- Balakrishnan K., Hsu F. J., Hafeman D. G., McConnell H. M. Monoclonal antibodies to a nitroxide lipid hapten. Biochim Biophys Acta. 1982 Sep 13;721(1):30–38. doi: 10.1016/0167-4889(82)90020-9. [DOI] [PubMed] [Google Scholar]
- Cuatrecasas P. Developing concepts in receptor research. Drug Intell Clin Pharm. 1983 May;17(5):357–366. doi: 10.1177/106002808301700507. [DOI] [PubMed] [Google Scholar]
- Davies D. R., Metzger H. Structural basis of antibody function. Annu Rev Immunol. 1983;1:87–117. doi: 10.1146/annurev.iy.01.040183.000511. [DOI] [PubMed] [Google Scholar]
- Hackenbrock C. R., Chazotte B., Gupte S. S. The random collision model and a critical assessment of diffusion and collision in mitochondrial electron transport. J Bioenerg Biomembr. 1986 Oct;18(5):331–368. doi: 10.1007/BF00743010. [DOI] [PubMed] [Google Scholar]
- Hart M. J., Kimura K., Nakanishi M. Selected positions of acyl chains are affected differently by antibody binding which results in decreased membrane fluidity. FEBS Lett. 1985 Oct 14;190(2):249–252. doi: 10.1016/0014-5793(85)81293-x. [DOI] [PubMed] [Google Scholar]
- Koppel D. E. Association dynamics and lateral transport in biological membranes. J Supramol Struct Cell Biochem. 1981;17(1):61–67. doi: 10.1002/jsscb.380170107. [DOI] [PubMed] [Google Scholar]
- McCloskey M., Poo M. M. Protein diffusion in cell membranes: some biological implications. Int Rev Cytol. 1984;87:19–81. doi: 10.1016/s0074-7696(08)62439-0. [DOI] [PubMed] [Google Scholar]
- McConnell H. M., Watts T. H., Weis R. M., Brian A. A. Supported planar membranes in studies of cell-cell recognition in the immune system. Biochim Biophys Acta. 1986 Jun 12;864(1):95–106. doi: 10.1016/0304-4157(86)90016-x. [DOI] [PubMed] [Google Scholar]
- Metzger H. The receptor on mast cells and related cells with high affinity for IgE. Contemp Top Mol Immunol. 1983;9:115–145. doi: 10.1007/978-1-4684-4517-6_4. [DOI] [PubMed] [Google Scholar]
- Nitta T., Saito-Taki T., Suzuki T. Phospholipase A2 activity of Fc gamma 2b receptors of thioglycollate-elicited murine peritoneal macrophages. J Leukoc Biol. 1984 Oct;36(4):493–504. doi: 10.1002/jlb.36.4.493. [DOI] [PubMed] [Google Scholar]
- O'Leary T. J. Concentration dependence of protein diffusion. Biophys J. 1987 Jul;52(1):137–139. doi: 10.1016/S0006-3495(87)83199-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Palmer A. G., 3rd, Thompson N. L. Molecular aggregation characterized by high order autocorrelation in fluorescence correlation spectroscopy. Biophys J. 1987 Aug;52(2):257–270. doi: 10.1016/S0006-3495(87)83213-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Place J. F., Sutherland R. M., Dähne C. Opto-electronic immunosensors: a review of optical immunoassay at continuous surfaces. Biosensors. 1985;1(4):321–353. doi: 10.1016/0265-928x(85)80004-3. [DOI] [PubMed] [Google Scholar]
- Sheetz M. P., Koppel D. E. Membrane damage caused by irradiation of fluorescent concanavalin A. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3314–3317. doi: 10.1073/pnas.76.7.3314. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith B. A., McConnell H. M. Determination of molecular motion in membranes using periodic pattern photobleaching. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2759–2763. doi: 10.1073/pnas.75.6.2759. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Subramaniam S., Seul M., McConnell H. M. Lateral diffusion of specific antibodies bound to lipid monolayers on alkylated substrates. Proc Natl Acad Sci U S A. 1986 Mar;83(5):1169–1173. doi: 10.1073/pnas.83.5.1169. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tamm L. K. Lateral diffusion and fluorescence microscope studies on a monoclonal antibody specifically bound to supported phospholipid bilayers. Biochemistry. 1988 Mar 8;27(5):1450–1457. doi: 10.1021/bi00405a009. [DOI] [PubMed] [Google Scholar]
- Tamm L. K., McConnell H. M. Supported phospholipid bilayers. Biophys J. 1985 Jan;47(1):105–113. doi: 10.1016/S0006-3495(85)83882-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thompson N. L., Brian A. A., McConnell H. M. Covalent linkage of a synthetic peptide to a fluorescent phospholipid and its incorporation into supported phospholipid monolayers. Biochim Biophys Acta. 1984 Apr 25;772(1):10–19. doi: 10.1016/0005-2736(84)90512-1. [DOI] [PubMed] [Google Scholar]
- Uzgiris E. E., Kornberg R. D. Two-dimensional crystallization technique for imaging macromolecules, with application to antigen--antibody--complement complexes. Nature. 1983 Jan 13;301(5896):125–129. doi: 10.1038/301125a0. [DOI] [PubMed] [Google Scholar]
- Uzgiris E. E. Supported phospholipid bilayers for two-dimensional protein crystallization. Biochem Biophys Res Commun. 1986 Jan 29;134(2):819–826. doi: 10.1016/s0006-291x(86)80494-6. [DOI] [PubMed] [Google Scholar]
- Yarden Y., Schlessinger J. Self-phosphorylation of epidermal growth factor receptor: evidence for a model of intermolecular allosteric activation. Biochemistry. 1987 Mar 10;26(5):1434–1442. doi: 10.1021/bi00379a034. [DOI] [PubMed] [Google Scholar]
- Young J. D., Unkeless J. C., Kaback H. R., Cohn Z. A. Macrophage membrane potential changes associated with gamma 2b/gamma 1 Fc receptor-ligand binding. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1357–1361. doi: 10.1073/pnas.80.5.1357. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Young J. D., Unkeless J. C., Kaback H. R., Cohn Z. A. Mouse macrophage Fc receptor for IgG gamma 2b/gamma 1 in artificial and plasma membrane vesicles functions as a ligand-dependent ionophore. Proc Natl Acad Sci U S A. 1983 Mar;80(6):1636–1640. doi: 10.1073/pnas.80.6.1636. [DOI] [PMC free article] [PubMed] [Google Scholar]